摘要
大肠杆菌的抗铜质粒中含有两个抗铜启动子,PpcoA和PpcoE,它们均含有copperbox。为了证实copper box在抗铜作用中的重要性以及研究抗铜启动子的特性,以质粒pUCD615为报告载体进行了这两个启动子不同长度片段的亚克隆。限制性酶切和DNA序列分析的结果表明,这些片段的亚克隆都是成功的;此外报告基因lux-荧光酶活性测定的结果表明,两个不含有copper box的Ppco short-lux亚克隆均不表达荧光酶活性,而其他启动子片段的亚克隆则都具有较高的酶活性,说明这两个亚克隆不具有启动子功能,初步证明在抗铜启动子中copper box是不可少的。
There were two promoters, PpcoA and PpcoE, in the copper resistant determinant from Escherichia coli. Either of them contains the copper box. In order to confirm the importance of copper box in the copper resistance promoters and to study their characteristics, several fragments of both promoters were subcloned using pUCD615 as reporter vector into its SmaI site. The results of restriction endonuclease digestion and the results of reporter gene lux-luciferase activities indicated that both of the Ppco short-lux fusions didn't show any luciferase activities, which indicated that these two fragments (Ppco short) couldn't act as promoters, and thus confirming the copper box was essential to copper resistance. If there were no copper box in the Ppco promoters, there would be no copper resistance in the E. coli.
出处
《微生物学报》
CAS
CSCD
北大核心
1997年第6期438-442,共5页
Acta Microbiologica Sinica
关键词
大肠杆菌
抗铜启动子
亚克隆
Escherichia coli, Copper resistant promoter, Subcloning, Copper box
